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在SARS-CoV的核酸检测方法中,由于普遍缺乏安全、稳定、特异的内对照,而不能对样品处理、反转录、扩增以及定量检测实施全程监控。构建的病毒样核蛋白颗粒内对照,能够对SARS-CoV临床检测实施监控。本文通过克隆1.7Kb大肠杆菌噬菌体MS2的装配蛋白和壳蛋白基因以及SARS冠状病毒经过突变产生的270bp内对照片断,并将这些基因连接到载体pTrc99a上表达,进行纯化、定量分析、RT-PCR检测和稳定性试验。获得SARS内对照病毒样核蛋白颗粒,在人血清和SM缓冲液中37℃稳定性可达到30天,能抵抗核糖核酸酶降解。将内对照颗粒加入临床样本中一同检测,能够对检测的全过程(样品处理、反转录和PCR)有效进行监控,与SARS-CoV没有交叉反应。研究表明制备的该病毒样核蛋白颗粒稳定、安全、可靠,可作为SARS冠状病毒RT-PCR检测、定量分析的有效内对照参考品。 相似文献
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JIN Xiang-qun ZHANG Jing-min XU Hui ZHOU Yan WANG Guang-shu ZHAO Yan-qiu ZHANG Han-qi 《高等学校化学研究》2006,22(2):185-188
Introduction Malignantdiseasesarecharacterizedbytheunreg ulatedgrowthoftransformedcells.Inrecentyears,dramaticinsightsintothemolecularmechanismsofthis phenomenonhavebeenachievedfrombasiccancerre search.Manycellularfunctionsareregulatedbychan gesingeneexpr… 相似文献
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Xinming Zhang Tianzhan Zhang Shuanli Guo Yang Zhang Rongtian Sheng Ruimeng Sun Lixia Chen Ruijuan Lv Yanfei Qi 《Molecules (Basel, Switzerland)》2020,25(24)
Fungal infections pose a serious threat to human health. Polyoxometalates (POMs) are metal–oxygen clusters with potential application in the control of microbial infections. Herein, the Ag3PW12O40 composites have been synthesized and verified by Fourier transform infrared (FT-IR) spectrum, transmission electron microscopy (TEM), scanning electron microscope (SEM), elemental analysis, and X-ray diffraction (XRD). The antifungal activities of Ag3PW12O40 were screened in 19 Candida species strains through the determination of minimum inhibitory concentration (MIC) by the microdilution checkerboard technique. The minimum inhibitory concentration (MIC50) values of Ag3PW12O40 are 2~32 μg/mL to the Candida species. The MIC80 value of Ag3PW12O40 to resistant clinical isolates C. albicans HL963 is 8 μg/mL, which is lower than the positive control, fluconazole (FLC). The mechanism against C. albicans HL963 results show that Ag3PW12O40 can decrease the ergosterol content. The expressions of ERG1, ERG7, and ERG11, which impact on the synthesis of ergosterol, are all prominently upregulated by Ag3PW12O40. It indicates that Ag3PW12O40 is a candidate in the development of new antifungal agents. 相似文献
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温敏核不育系水稻株1S及其矮秆突变体SV14茎的蛋白质组学比较 总被引:1,自引:0,他引:1
采用双向凝胶电泳对温敏核不育水稻株1S和其矮秆突变体SV14的茎(穗颈下第1节和第2节)蛋白进行了分离, 通过银染显色, 获得了分辨率和重复性较好的双向电泳图谱. 选取了26个蛋白质点采用MALDI-TOF-MS进行肽质谱指纹图分析, 最终有12个蛋白质点得到了可靠鉴定. 其中在SV14中相对于株1S上调的仅有OSJNBa0039C07.13 蛋白, 其它蛋白均表现为下调. 这些差异蛋白按照功能可分为4类: (1) 能量代谢相关蛋白; (2) 次生代谢相关蛋白; (3) 调控蛋白; (4) 未知蛋白. 对光合系统Ⅱ氧延伸复合物蛋白质前体2, 果糖二磷酸醛缩酶, UDP-葡糖醛酸脱羧酶对应的基因进行了半定量RT-PCR分析, 发现这几个基因与蛋白质的表达不一致, 可能是RNA发生了翻译后修饰而减少了蛋白表达量的结果. 这些差异蛋白很可能与水稻矮化有关, 为水稻矮秆基因的寻找提供了另一个有效途径. 相似文献
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In this study, biostimulation technology was used for bioremediation of nitrobenzene-contaminated groundwater by adding a mixture of lactose and phosphate, peptone, and beef extract. During the process of biostimulation, the remediation effectiveness, microbial dehydrogenase activities and microbial densities were investigated; the varieties of microbial community structure and composition were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE) technique and the relative abundances of nitrobenzene-degrading gene(nbzA) were determined by fluorescence quantitative real-time PCR(RT-PCR). Findings show that the removal rate of nitrobenzene in groundwater could reach about 60% by biostimulation with lactose and phosphate, 70% with peptone and 68% with beef extract. The microbial dehydrogenase activities and microbial densities were all improved obviously via biostimulation. The results of PCR-DGGE show that the microbial diversities were improved, and more than ten kinds of dominant microorganisms were detected after biostimulation. RT-PCR results show that the relative abundances of nbzA gene of microbes in groundwater were increased significantly, which indicated that biostimulation actually enhanced the growth of nitrobenzene-degrading bacteria. Therefore, biostimulation is a cost-effective and feasible bioremediation technique for nitrobenzene-contaminated groundwater. 相似文献